Unlocking the Secrets of Adipose Tissue: How an Obesity-Associated Secretome Promotes Osteoblast Dedifferentiation via TGF-ß1 Signaling, Paving the Path to an Adipogenic Phenotype.

BACKGROUND: Obesity poses a significant global health challenge, given its association with the excessive accumulation of adipose tissue (AT) and various systemic disruptions. Within the adipose microenvironment, expansion and enrichment with immune cells trigger the release of inflammatory mediators and growth factors, which can disrupt tissues, including bones. While obesity's contribution to bone loss is well established, the direct impact of obese AT on osteoblast maturation remains uncertain. This study aimed to explore the influence of the secretomes from obese and lean AT on osteoblast differentiation and activity. METHODS: SAOS-2 cells were exposed to the secretomes obtained by culturing human subcutaneous AT from individuals with obesity (OATS) or lean patients, and their effects on osteoblasts were evaluated. RESULTS: In the presence of the OATS, mature osteoblasts underwent dedifferentiation, showing an increased proliferation accompanied by a morphological shift towards a mesenchymal phenotype, with detrimental effects on osteogenic markers and the calcification capacity. Concurrently, the OATS promoted the expression of mesenchymal and adipogenic markers, inducing the formation of cytoplasmic lipid droplets in SAOS-2 cells exposed to an adipogenic differentiation medium. Additionally, TGF-ß1 emerged as a key mediator of these effects, as the OATS was enriched with this growth factor. CONCLUSIONS: Our findings demonstrate that obese subcutaneous AT promotes the dedifferentiation of osteoblasts and increases the adipogenic profile in these cells.

Correlation between sporogenesis and lipopeptide production in Paenibacillus elgii.

Pelgipeptins, tridecaptins, and elgicins are among the antimicrobials produced by Paenibacillus elgii. Growth in complex media is commonly applied to obtain lipopeptides from culture's supernatant, but it requires further purification. This study aimed to improve the yield of pelgipeptins and tridecaptins using chemically defined media. The kinetics of antimicrobial lipopeptide yield in chemically defined media were evaluated in P. elgii AC13. Pelgipeptins were detected in the supernatant and the culture pellet, but tridecaptins were mainly associated with cell debris or endospores. We investigated whether removing Ca2+ would impair P. elgii sporogenesis, consequently improving the yield of tridecaptin. The kinetics of both lipopeptides in the presence and absence of Ca2+ were quantitatively and qualitatively evaluated and further correlated with the cell cycle. The impairment of P. elgii AC13 sporogenesis had no effect on tridecaptin production, which remained undetected in the supernatant of the culture. On the other hand, the yield of pelgipeptin in a Ca2+-free medium increased. We showed for the first time that the removal of Ca2+ interrupted the sporogenesis in P. elgii and improved the yield of pelgipeptins. However, Ca2+ absence had no effect on tridecaptin yield, which is possibly degraded or associated with other cell debris components.

VL-HIV co-infection with Leishmania containing skin lesions resembling para-kala-azar dermal leishmaniasis.

Leishmaniases are a group of neglected vector-borne infectious diseases that are among the six priority endemic diseases worldwide. Visceral leishmaniasis (VL) is the most severe clinical manifestation, characterized by systemic and chronic visceral involvement and high mortality in immunosuppressed and untreated patients. VL can be complicated into post-kala-azar dermal leishmaniasis (PKDL), and when dermatologic disorders occur simultaneously with active VL, an intermediate clinical form called para-kala-azar dermal leishmaniasis (para-KDL) occurs. This clinical form is of great epidemiological relevance, as humans act as a source of infection for vectorial transmission. In the Americas, Brazil is among the seven countries responsible for more than 90% of VL cases, though reports of PKDL and para-KDL are rare. This paper presents three cases of VL-HIV co-infection with Leishmania-containing skin lesions resembling para-kala-azar dermal leishmaniasis. The cases were investigated by the team from the Infectious Diseases Department of University Hospital (HUMAP/UFMS) in Mato Grosso do Sul, Brazil. The three patients exhibited skin lesions where amastigote forms of L. (L.) infantum were identified. All cases exhibited similar clinical manifestations of para-KDL, including fever, hepatosplenomegaly, pancytopenia, and disseminated skin lesions. The study described the prevalence of comorbidities, the incidence of VL relapse, and the therapeutic regimen in relation to the outcomes. The study underscores the importance of follow-up and secondary prophylaxis in patients with VL, which are essential for the efficacy of the treatment. Furthermore, the study provides insight into the potential epidemiological profile of para-KDL cases in Brazil, which contributes to the development of more efficient clinical management strategies for patients.

In silico optimization of analogs derived pro-adrenomedullin peptide to evaluate antimicrobial potential.

Diverse computational approaches have been widely used to assist in designing antimicrobial peptides with enhanced activities. This tactic has also been used to address the need for new treatment alternatives to combat resistant bacterial infections. Herein, we have designed eight variants from a natural peptide, pro-adrenomedullin N-terminal 20 peptide (PAMP), using an in silico pattern insertion approach, the Joker algorithm. All the variants show an α-helical conformation, but with differences in the helix percentages according to circular dichroism (CD) results. We found that the C-terminal portion of PAMP may be relevant for its antimicrobial activities, as revealed by the molecular dynamics, CD, and antibacterial results. The analogs showed variable antibacterial potential, but most were not cytotoxic. Nevertheless, PAMP2 exhibited the most potent activities against human and animal-isolated bacteria, showing cytotoxicity only at a substantially higher concentration than its minimal inhibitory concentration (MIC). Our results suggest that the enhanced activity in the profile of PAMP2 may be related to their particular physicochemical properties, along with the adoption of an amphipathic α-helical arrangement with the conserved C-terminus portion. Finally, the peptides designed in this study can constitute scaffolds for the design of improved sequences.

Pelgipeptins, a Nonribosomal Lipopeptide Family, Show Larvicidal Activity against Vectors Transmitting Viruses.

Aedes aegypti and Culex quinquefasciatus are vectors of numerous diseases of worldwide public importance, such as arboviruses and filariasis. The main strategy for controlling these vectors is the use of chemicals, which can induce the appearance of resistant insects. The use of Bacillus thuringiensis (Bt) and Lysinibacillus sphaericus (Ls) with larvicidal activity against arboviral-transmitting insects has been successful in many studies. In contrast, the use and knowledge of peptides with insecticidal activity are so far scarce. In this work, 25 peptides and 5 strains of each bacterial species were prospected individually or together regarding their insecticidal activity. Initially, in vitro assays of cellular cytotoxicity of the peptides against SF21 cells of Spodoptera frugiperda were performed. The peptides Polybia-MPII and pelgipeptin caused 69 and 60% of cell mortality, respectively, at the concentration of 10 µM. Thus, they were evaluated in vivo against second-stage larvae of the two Culicidae. However, in the in vivo bioassays, only pelgipeptin showed larvicidal mortality against both larvae (LC50 6.40 µM against A. aegypti, and LC50 1.22 µM against C. quinquefasciatus). The toxin-producing bacterial strain that showed the lowest LC50 against A. aegypti was Bt S8 (LC50 = 0.71 ng/mL) and against C. quinquefasciatus, it was Ls S260 (LC50 = 2.32 ng/mL). So, the synergistic activity between the association of the bacterial toxins and pelgipeptin was evaluated. A synergic effect of pelgipeptin was observed with Ls strain S260 against C. quinquefasciatus. Our results demonstrate the possibility of synergistic or individual use of both biologically active larvicides against C. quinquefasciatus and A. aegypti.

Promoção de saúde e qualidade de vida dos universitários em tempos de pandemia: a experiência de um curso de extensão

Objetivos: Este artigo apresenta o relato da experiência do curso/workshop RENASCERES®: saúde e qualidade de vida dos universitários em tempos de pandemia", realizado em uma universidade federal no Estado de Minas Gerais (MG), que teve o objetivo de criar espaço de diálogo, acolhimento, escuta e de construção de alternativas mediante demandas discentes, decorrentes do período de pandemia de COVID-19. Objetivou também fomentar propostas para a ampliação das condições de saúde e da qualidade de vida dos universitários e de suas famílias. Métodos: O curso foi estruturado em cinco encontros virtuais, com formato de workshop, fundamentados na metodologia dialógica e no método de Círculo de Cultura, articulando o conteúdo teórico aliado às atividades práticas e ao cotidiano dos participantes. Resultados: Participaram do curso 77 pessoas, incluindo universitários de diferentes cursos de graduação e profissionais. As reflexões sobre saúde e qualidade de vida e o atual contexto de pandemia, bem como as atividades práticas, geraram espaço de acolhimento, escuta-ativa, cuidado, acesso e troca de informações/novos conhecimentos. Conclusões: O curso/workshop possibilitou o reconhecimento da necessidade de investimentos em Promoção de saúde e qualidade de vida pelos participantes. Constatou-se, nesta atividade, a essencialidade do Método RENASCERES® como alternativa para dar suporte aos participantes nos cuidados em saúde, mudanças em suas rotinas e pilar para que as pessoas possam alcançar estilos de vida saudáveis.Palavras-chave: Educação para a saúde; Promoção da saúde; Qualidade de vida; Covid-19; Saúde Coletiva; Comunicação em saúde.

A novel family of non-secreted tridecaptin lipopeptide produced by Paenibacillus elgii.

Bacteria from the genus Paenibacillus make a variety of antimicrobial compounds, including lipopeptides produced by a non-ribosomal synthesis mechanism (NRPS). In the present study, we show the genomic and phenotypical characterization of Paenibacillus elgii AC13 which makes three groups of small molecules: the antimicrobial pelgipeptins and two other families of peptides that have not been described in P. elgii. A family of lipopeptides with [M + H]+ 1664, 1678, 1702, and 1717 m/z was purified from the culture cell fraction. Partial characterization revealed that they are similar to tridecaptin from P. terrae. However, they present amino acid chain modifications in positions 3, 7, and 10. These new variants were named tridecaptin G1, G2, G3, and G4. Furthermore, a gene cluster was identified in P. elgii AC13 genome, revealing high similarity to the tridecaptin-NRPS gene cluster from P. terrae. Tridecaptin G1 and G2 showed in vitro antimicrobial activity against Escherichia coli, Klebsiella pneumonia (including a multidrug-resistant strain), Staphylococcus aureus, and Candida albicans. Tri G3 did not show antimicrobial activity against S. aureus and C. albicans at all tested concentrations. An intriguing feature of this family of lipopeptides is that it was only observed in the cell fraction of the P. elgii AC13 culture, which could be a result of the amino acid sequence modifications presented in these variants.

Fifteen years of enzyme replacement therapy for mucopolysaccharidosis type VI (Maroteaux-Lamy syndrome): a case report.

BACKGROUND: Mucopolysaccharidosis VI, or Maroteaux-Lamy disease, is an autosomal recessive disease characterized by deficiency of the enzyme arylsulfatase B in the lysosomal catabolism of glycosaminoglycans. Due to reduced (or even null) enzyme activity, glycosaminoglycans (mainly dermatan sulfate) accumulates, leading to a multisystemic disease. Mucopolysaccharidosis VI induces reduced growth, coarse face, audiovisual deficits, osteoarticular deformities, and cardiorespiratory issues, hampering the quality of life of the patient. Enzyme replacement therapy with galsulfase (Naglazyme, BioMarin Pharmaceuticals Inc., USA) is the specific treatment for this condition. Although studies have shown that enzyme replacement therapy slows the progression of the disease, the effects of long-term enzyme replacement therapy remain poorly understood. CASE PRESENTATION: A 29-year-old, Caucasian, male patient diagnosed with mucopolysaccharidosis VI was treated with enzyme replacement therapy for over 15 years. Enzyme replacement therapy was initiated when patient was 13 years old. The patient evolved multiplex dysostosis, carpal tunnel syndrome, thickened mitral valve, and hearing and visual loss. CONCLUSIONS: Although enzyme replacement therapy did not prevent the main signs of mucopolysaccharidosis VI, it slowed their progression. Additionally, enzyme replacement therapy was associated with a longer survival compared with the untreated affected sibling. Taken together, the results indicate that enzyme replacement therapy positively modified the course of the disease.

Co-occurrence of linear and cyclic pelgipeptins in broth cultures of Paenibacillus elgii AC13.

Paenibacillus elgii AC13 produces antimicrobial lipopeptides of agricultural and pharmaceutical importance. It secretes four cyclic lipopeptides named pelgipeptins, previously characterized in P. elgii B69. These lipopeptides result from the expression of a nonribosomal peptide gene cluster. P. elgii AC13 also produced two linear lipopeptides with ratios of [M + H] + 1105 and 1119 m/z. These compounds were previously observed in Paenibacillus sp. strain OSY-N, but due to purification difficulties, their characterization was executed using synthetically produced linear pelgipeptins. In the present study, purification was achieved from the supernatants of cultures from three complex media by high-performance liquid chromatography. The partial characterization of linear pelgipeptins revealed the similar antimicrobial activity and cytotoxicity of their synthetically produced counterparts, known as paenipeptins. Cyclic forms were highly stable to changes in pH, temperature, and organic extraction with n-butanol as shown by mass spectrometry (MALDI-TOF); therefore, these steps did not cause the hydrolysis of pelgipeptins. A low-activity thioesterase could also generate the linear isoforms observed; this enzyme catalyzes the cyclization process and is coded in the same gene cluster. Alternatively, the cyclic forms were hydrolyzed by an unknown protease produced during growth in the complex medium used in the present study. Although culture conditions are known to produce pelgipeptins with different yields and amino acid compositions, the occurrence of linear and cyclic forms simultaneously has not yet been reported. A mixture of cyclic and linear pelgipeptins presents a potential advantage of the higher antimicrobial activity of cyclic forms combined with the lower cytotoxicity of linear isoforms.

Adipose Tissue-Derived Extracellular Vesicles and the Tumor Microenvironment: Revisiting the Hallmarks of Cancer.

Extracellular vesicles (EVs) are crucial elements that sustain the communication between tumor cells and their microenvironment, and have emerged as a widespread mechanism of tumor formation and metastasis. In obesity, the adipose tissue becomes hypertrophic and hyperplastic, triggering increased production of pro-inflammatory adipokines, such as tumor necrosis factor α, interleukin 6, interleukin 1, and leptin. Furthermore, obese adipose tissue undergoes dysregulation in the cargo content of the released EVs, resulting in an increased content of pro-inflammatory proteins, fatty acids, and oncogenic microRNAs. These alterations drive obesity-associated inflammatory responses both locally and systemically. After being ignored for a long time, adipose tissues have recently received considerable attention as a major player in tumor microenvironment-linked obesity and cancer. The role of adipose tissue in the establishment and progression of cancer is reinforced by its high plasticity and inflammatory content. Such a relationship may be established by direct contact between adipocytes and cancer cells within the microenvironment or systemically, via EV-mediated cell-to-cell communication. Here, we highlight cues evidencing the influence of adipose tissue-derived EVs on the hallmarks of cancer, which are critical for tumor malignancy.

Obese adipose tissue extracellular vesicles raise breast cancer cell malignancy.

Obesity is a chronic low-grade inflammatory condition that strongly impacts breast cancer. Aside from inflammatory mediators, obese adipose tissue (AT) secretes high amounts of extracellular vesicles (EVs), which are capable of transferring molecules to target cells and promoting cell-to-cell communication. Here, we investigated how soluble mediators and EVs secreted by human obese AT influence MCF-7 and MDA-MB-231 mammary adenocarcinoma cell lines by modulating cell proliferation, migration, invasion, and signaling pathways. Both cell lineages were stimulated with conditioned media (CM) or EVs obtained from cultures of AT explants collected from lean or obese individuals who underwent plastic or bariatric surgeries, respectively. EVs derived from obese AT increased the proliferative potential of both cell lines and further potentiated the migratory and invasive properties of MDA-MB-231 cells. The proliferative effects of CM and EVs on MCF-7 cells were dependent on ERK/MAPK pathway activation, while the migration and invasiveness of MDA-MB-231 cells were dependent on PI3K/AKT pathway activation. Furthermore, CM derived from obese AT potentiated the pro-angiogenic effect of MDA-MB-231 on endothelial cells. We also detected that EVs derived from obese AT were enriched in leptin and bioactive matrix metallopeptidase 9 (MMP9), and stimulation of MDA-MD-231 cells with those EVs or CM derived from obese AT potentiated the release of MMP9 by those cells. Our data indicate that obese AT secretes molecules and EVs with pro-tumoral activities capable of increasing breast cancer cell malignancy and provide strong evidence of the key role of AT-derived EV signaling in the tumor microenvironment.

Microparticles derived from obese adipose tissue elicit a pro-inflammatory phenotype of CD16+, CCR5+ and TLR8+ monocytes.

Macrophage infiltration into adipose tissue (AT) is a hallmark of the chronic inflammatory response in obesity and is supported by an intense monocyte migration towards AT. Although it has been detected an increased proportion of circulating CD16+ monocyte subsets in obese subjects, the mechanisms underlying this effect and the contribution of these cells to the inflamed profile of obese AT are still poorly understood. We investigated whether factors secreted by human obese omental AT could polarize monocytes to CD16+ enriched phenotype, and how these changes could modify their migratory capacity towards adipose tissue itself. We show that explants of human obese omental AT, obtained during bariatric surgery, released higher levels of MIP1-α, TNFα, leptin and also VEGF, together with increasing amounts of microparticles (MP), when compared to explants of lean subcutaneous AT. A higher content of circulating MP derived from preadipocytes and leukocytes was also detected in plasma of obese subjects. Conditioned media or MP released from obese omental AT increased CD16 and CCR5 expression on CD14+CD16- monocytes and augmented their migratory capacity towards the conditioned media from obese omental AT, itself. This effect was inhibited when MIP1-α was neutralized. Additionally, we demonstrate that MP derived from obese omental AT carry and transfer TLR8 to monocytes, thus triggering an increase in CD16 expression in those cells. Our data shows a positive feedback loop between blood monocytes and obese omental AT, which releases chemotactic mediators and TLR8-enriched MP, thus inducing an up-regulation of CD16+ monocytes, favoring leukocyte infiltration in the obese omental AT.

Empregabilidade do psicólogo: análise da percepção de alunos, ex-alunos, professores universitários e selecionadores / Employability of psychologists: analysis of the perception of undergraduate students, former students, professors and selectors

O objetivo deste estudo foi analisar se o curso de Psicologia de uma universidade particular, segundo a ótica de professores, alunos, ex-alunos e selecionadores possibilita o desenvolvimento de competências necessárias para a conquista de empregabilidade. Foram realizadas análise descritiva de questionários aplicados em 82 alunos e ex-alunos, análise descritiva e de conteúdo de 43 entrevistas com alunos, ex-alunos (que atuam e que não atuam como psicólogos), professores, selecionadores e com a coordenadora do Programa de apoio à Carreira. Os dados apontam que 50% dos exalunos estão inseridos no mercado de trabalho, destes 40% percebem que adquiriram competências através da graduação. Os resultados sugerem que as IES façam discussões sobre empregabilidade envolvendo alunos, ex-alunos, professores e selecionadores.

The objective of this study was to analyze if the psychology course in a private university from the perspective of teachers, students, alumni and selectors can enable the development of skills needed for achieving employability. Descriptive analysis were made of questionnaires in 82 students and former students, descriptive and content analysis of 43 interviews with students, former students (who work and do not work as psychologists), teachers, selectors and the coordinator of the support program career. The data indicate that 50% of former students are in the job market, of these 40% realise that have acquired skills trought graduation. The results suggest that higher education institutions need to hold discussions on employability involving students, alumni, teachers and selectors.

Empregabilidade do psicólogo: análise da percepção de alunos, ex-alunos, professores universitários e selecionadores / Employability of psychologists: analysis of the perception of undergraduate students, former students, professors and selectors

O objetivo deste estudo foi analisar se o curso de Psicologia de uma universidade particular, segundo a ótica de professores, alunos, ex-alunos e selecionadores possibilita o desenvolvimento de competências necessárias para a conquista de empregabilidade. Foram realizadas análise descritiva de questionários aplicados em 82 alunos e ex-alunos, análise descritiva e de conteúdo de 43 entrevistas com alunos, ex-alunos (que atuam e que não atuam como psicólogos), professores, selecionadores e com a coordenadora do Programa de apoio à Carreira. Os dados apontam que 50% dos exalunos estão inseridos no mercado de trabalho, destes 40% percebem que adquiriram competências através da graduação. Os resultados sugerem que as IES façam discussões sobre empregabilidade envolvendo alunos, ex-alunos, professores e selecionadores. (AU)

The objective of this study was to analyze if the psychology course in a private university from the perspective of teachers, students, alumni and selectors can enable the development of skills needed for achieving employability. Descriptive analysis were made of questionnaires in 82 students and former students, descriptive and content analysis of 43 interviews with students, former students (who work and do not work as psychologists), teachers, selectors and the coordinator of the support program career. The data indicate that 50% of former students are in the job market, of these 40% realise that have acquired skills trought graduation. The results suggest that higher education institutions need to hold discussions on employability involving students, alumni, teachers and selectors. (AU)